Organic non-sugar compounds for protection of biologically active molecules and conjugate labels and methods of use thereof

ABSTRACT

Provided are methods comprising the use of non-sugar organic compatible solutes for protection and preservation of the activity of biologically active molecules and conjugate labels. The methods are particularly adaptable for use in conjunction with immunoassays, such as for example, immunochromatographic test assays and may be incorporated into any test methodology wherein a dry test strip is used as a carrier for depositing, mobilizeable and/or immobilized biologically active molecules and/or conjugate labels.

RELATED APPLICATIONS

This application claims the benefit of priority to PCT/US2006/024955filed Jun. 27, 2006, which claims priority to U.S. Provisional PatentApplication 60/694,604, filed Jun. 27, 2005, which applications arehereby incorporated by reference in their entirety.

BACKGROUND OF THE INVENTION

A variety of buffers and stabilizers for use in conjunction withproteins are commercially available. These solvents and buffers can beused in a variety of procedures and tests including immunoassays. Manyof the available buffers and stabilizers become ineffective inprotecting and preserving the activity of biologically active moleculesand conjugate labels under the dry conditions of immunochromatographictest assays. Some immunochromatographic test assays have a very shortshelf-life and oftentimes lose effectiveness or become totallyineffective in a short period of time due to the degradation of theactivity of the molecules during storage in dry conditions. While therehas been a trend to produce more effective and longer lastingimmunochromatographic test assays, further improvements foreffectiveness and preservation of activity of active molecules andconjugate labels are desirable.

SUMMARY OF THE INVENTION

Provided are methods of using non-sugar organic compatible solutes forprotection and preservation of the activity of biologically activemolecules and conjugate labels. We have found that the activity ofbiologically active molecules deposited on dry carriers ofimmunochromatographic test (ICT) assays can be preserved, protected andkept preserved for longer durations of time when deposited together withone or more non-sugar organic compatible solutes. Non-sugar organiccompatible solutes may be used in conjunction with immunoassays, such asfor example ICT assays, but also in any application wherein theprotection and preservation of the activity of biologically activemolecules and conjugate labels is desired, for example, in solutionphase, in dry state, or in reconstituted state. The non-sugar organiccompatible solutes may be combined with any test methodology wherein adry test strip is used as a carrier for depositing mobilizable and/orimmobilized biologically active molecules and/or conjugate labels.

DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts the stabilizing effect that a combination of 2% w/v ofecotine, L-histidine, taurine and L-threonine has on conjugate paddrying buffer without sucrose for a malaria ICT device. Equivalentsensitivity was observed for conjugate pad stored at both 4° C. and 55°C. despite the absence of sucrose in the drying pad buffer formulation.

FIG. 2 illustrates that 4% w/v of L-Histidine alone does not conferstability to conjugate pad drying buffer without sucrose for a malariaICT device.

DETAILED DESCRIPTION OF THE INVENTION 1. Introduction

The activity of biologically active molecules can generally bepreserved, protected, and kept preserved for longer durations of timewhen deposited together with one or more non-sugar organic compatiblesolutes, particularly when deposited on dry carriers ofimmunochromatographic test assays.

In one aspect, provided is a method comprising:

a) depositing one or more biologically active molecules onto a surface;

b) depositing one or more non-sugar organic compatible solutes onto thesurface; and

c) drying the deposited one or more biologically active molecules andthe deposited one or more non-sugar organic compatible solutes on thesurface; wherein the one or more non-sugar organic compatible solutesprotects and preserves the activity of the one or more biologicallyactive molecules in the dried and reconstituted state.

In another aspect, provided is a method comprising:

a) providing a sample;

b) providing a test strip with at least a portion comprising one or morebiologically active molecules and one or more non-sugar organiccompatible solutes, wherein the one or more non-sugar organic compatiblesolutes protects and preserves the activity of the one or morebiologically active molecules in the dried and reconstituted state.

c) contacting the sample with the test strip; and

d) detecting the presence of or measuring the amount of an analyte ofinterest in the sample.

In another aspect, provided is a method comprising:

a) providing one or more biologically active molecules; and

b) providing one or more non-sugar organic compatible solutes; whereinthe one or more non-sugar organic compatible solutes preserves theactivity of the one or more biologically active molecules.

Further objectives and advantages of the present invention will becomeapparent as the description proceeds. To gain a full appreciation of thescope of the present invention, it will be further recognized thatvarious aspects of the present invention can be combined to makedesirable embodiments of the invention.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Where a term is provided in thesingular, the inventor also contemplates the plural of that term. Thenomenclature used herein and the procedures described below are thosewell known and commonly employed in the art.

The term “amino acid” is intended to embrace all molecules, whethernatural or synthetic, which include both an amino functionality and anacid functionality and capable of being included in a polymer ofnaturally-occurring amino acids. Exemplary amino acids includenaturally-occurring amino acids; analogs, derivatives and congenersthereof; amino acid analogs having variant side chains; and allstereoisomers of any of any of the foregoing. The names of the naturalamino acids are abbreviated herein in accordance with therecommendations of IUPAC-IUB.

The term “antibody” refers to an immunoglobulin, derivatives thereofwhich maintain specific binding ability, and proteins having a bindingdomain which is homologous or largely homologous to an immunoglobulinbinding domain. These proteins may be derived from natural sources, orpartly or wholly synthetically produced. An antibody may be monoclonalor polyclonal. The antibody may be a member of any immunoglobulin class,including any of the human classes: IgG, IgM, IgA, IgD, and IgE. Inexemplary embodiments, antibodies used with the methods and compositionsdescribed herein are derivatives of the IgG class.

The term “antibody fragment” refers to any derivative of an antibodywhich is less than full-length. In exemplary embodiments, the antibodyfragment retains at least a significant portion of the full-lengthantibody's specific binding ability. Examples of antibody fragmentsinclude, but are not limited to, Fab, Fab′, F(ab′)2, scFv, Fv, dsFvdiabody, and Fd fragments. The antibody fragment may be produced by anymeans. For instance, the antibody fragment may be enzymatically orchemically produced by fragmentation of an intact antibody, it may berecombinantly produced from a gene encoding the partial antibodysequence, or it may be wholly or partially synthetically produced. Theantibody fragment may optionally be a single chain antibody fragment.Alternatively, the fragment may comprise multiple chains which arelinked together, for instance, by disulfide linkages. The fragment mayalso optionally be a multimolecular complex. A functional antibodyfragment will typically comprise at least about 50 amino acids and moretypically will comprise at least about 200 amino acids.

The term “biologically active molecule” refers to any active organicmolecule that is part of a living organism, which may be natural,synthetic, or a combination thereof. The terms “comprise” and“comprising” is used in the inclusive, open sense, meaning thatadditional elements may be included.

The term “including” is used herein to mean “including but not limitedto”. “Including” and “including but not limited to” are usedinterchangeably.

The terms “label” or “labeled” refer to incorporation or attachment,optionally covalently or non-covalently, of a detectable marker into amolecule, such as a polypeptide, e.g., an antibody. Various methods oflabeling polypeptides are known in the art and may be used. Examples oflabels for polypeptides include, but are not limited to, the following:radioisotopes, fluorescent labels, heavy atoms, enzymatic labels orreporter genes, chemiluminescent groups, biotinyl groups, predeterminedpolypeptide epitopes recognized by a secondary reporter (e.g., leucinezipper pair sequences, binding sites for secondary antibodies, metalbinding domains, epitope tags). In some embodiments, labels are attachedby spacer arms of various lengths to reduce potential steric hindrance.

The term “test strip” refers to a chromatographic-like medium upon whichan assay may be performed.

The term “sample” refers to any fluid sample potentially containing ananalyte.

2. Methods for Preserving Activity of Biologically Active Molecules inthe Dried and Reconstituted State

Provided is the use of non-sugar organic compatible solutes forprotection and preservation of the activity of biologically activemolecules and conjugate labels. The methods described below areadaptable for use with an immunochromatographic test strip having aporous carrier such as a nitrocellulose test strip, which is well-knownin the art.

Provided is a method for preserving the activity of biologically activemolecules in the dried and reconstituted state comprising depositing oneor more biologically active molecules onto a surface. The one or morebiologically active molecules can be deposited onto the surface usingany suitable method. The one or more biologically active molecules canbe any suitable biologically active molecule, such as, for example, abiologically active molecule that is capable of specifically binding oneor more analytes of interest. Suitable molecules of the presentinvention include, for example, polyclonal or monoclonal antibodies orantibody fragments. The surface can be any type of surface suitable forperforming immunoassays, such as, for example, a nitrocellulose teststrip.

In certain embodiments, the surface or test strip is part of a lateralflow device. In a lateral flow device, a sample containing a targetanalyte of interest is added to an application well in the lateral flowdevice. The test sample flows along a test strip or surface toward azone containing a mobilizable binding reagent disposed in the zone in adry state, e.g., in a pad. When the sample front reaches this zone, themobilizable labeled binding reagent is released from the pad and thenallowed to interact with target analyte that may be present in the testsample. Target analyte in the sample binds to the mobilizable bindingagent and flow continues toward a detection zone. The detection zonecomprises an immobilized unlabelled specific binding reagent, forexample, on a pad, for binding to the analyte/mobilizable binding agentcomplexes. The presence of analyte in the sample is determined byobserving the extent (if any) to which the labeled binding agent/analytecomplex becomes bound in the detection zone. Results can be assessedvisually by colorometric means with the intensity of color beingdirectly proportional to target analyte concentration within the testsample. In one embodiment, the labeled reagent, the analyte (if present)and the immobilized unlabelled specific binding agent cooperate togetherin a “sandwich” reaction. This results in the labeled reagent beingbound in the detection zone if analyte is present in the sample. The twobinding reagents must have specificities for different epitopes on theanalyte.

An exemplary lateral flow device of this type is disclosed in Davis, etal., U.S. Pat. No. 6,352,862, which is incorporated by reference hereinin its entirety.

The method may also comprise depositing one or more non-sugar organiccompatible solutes onto a surface. The deposition of the one or morebiologically active molecules and the one or more non-sugar organiccompatible solutes onto the surface of a carrier or surface can be donein any suitable manner such as concurrent or separate deposition. Theone or more non-sugar organic compatible solutes of the presentinvention can be any non-sugar organic compatible solutes suitable forthe purpose of the invention, namely, effective to preserve, andprotect, the activity of the one or more biologically active moleculesfor a longer duration of time than would be possible without thepresence of the one or more non-sugar organic compatible solutes. Theone or more non-sugar organic compatible solutes of the presentinvention include, for example, amino acids such as glycine, alanine,serine, proline, sodium glutamate, lysine, aminobutric acid, taurine,amines such as betaine, sarcosine, trimethylamine N-oxide, or osmolytesor other compatible solutes such as 2-naphthol, ectoine, hydroxyectoine,glycerine, deoxyribonucleic acid, adenosine monophosphate, PCR primers,spermine, spermidine, Brij58, polyethyleneimine, polyhystidine,polythymadilic acid, acetyl-L-carnitine, L-carline, hypotaurine, andferulic acid.

The methods may also comprise drying the deposited one or morebiologically active molecules and the deposited one or more non-sugarorganic compatible solutes on a surface. Any suitable drying methodologyknown in the art may be used for drying the deposited one or morebiologically active molecules and the one or more non-sugar organiccompatible solutes.

An important aspect of the methods is the ability of the one or morenon-sugar organic compatible solutes to protect and preserve theactivity of the one or more biologically active molecules in the driedand reconstituted state.

3. Methods for Detecting the Presence of or Measuring the Amount of anAnalyte of Interest in a Sample

Further provided is the use of non-sugar organic compatible solutes forprotection and preservation of the activity of biologically activemolecules and conjugate labels and methods thereof. The methods providedherein are adaptable for use with an immunochromatographic test striphaving a porous carrier such as a nitrocellulose test strip, forexample, a lateral flow device, which are numerous and vary intechniques and are well known in the art as described in detail in theprevious section.

By way of example, generally, an immunoassay device for determining thepresence or amount of an analyte of interest in a sample includes asample application member, which is in liquid communication with aconjugate pad, which is in liquid communication with a nitrocellulosetest strip having a test result zone and a control zone. The immunoassaycan also include a distal sink at the end opposite to the sampleapplication pad to absorb any excess liquid after testing has run tocompletion.

The sample application pad is a porous pad able to absorb the sample tobe tested and transfer the absorbed sample to the conjugate pad bycapillary action. The conjugate pad includes one or more dried labeledmolecules or reagents, such as antibodies, capable of specificallybinding to the one or more analytes of interest forming aanalyte-labeled reagent complex. The conjugate pad may also include oneor more stabilizing compounds that are able to induce thermal stabilityand also stability as to conditions imposed by humidity and temperature.The conjugate pad is a porous pad able to absorb the transferred samplefrom the sample application pad and transfer the sample to thenitrocellulose strip by capillary action. The nitrocellulose strip isable to absorb the sample from the conjugate pad and transfer the sampleby capillary action downstream to the test result zone and the controlzone. The test result zone of the immunoassay device includes one ormore immobilized molecules or reagents, such as antibodies, capable ofspecifically binding to the one or more analytes of interest or anyportion of the analyte-labeled reagent complex. The control zone of theimmunoassay device may include one or more immobilized molecules orreagents, such as antibodies, capable of specifically binding to the oneor more labeled reagent.

When a liquid test sample is applied to the sample application pad ofthe device, the sample travels through the sample application pad, theconjugate pad, and nitrocellulose strip by capillary action. When thesample travels through the conjugate pad, the sample solublizes thedried labeled molecule or reagent, and if the analyte of interest ispresent in the sample, the solublized labeled molecule or reagent bindsthe analyte of interest forming an analyte-labeled reagent complex,otherwise, if the analyte of interest is not present in the sample, nocomplex is formed. The analyte-labeled reagent complex in the case of apositive test, or the labeled reagent alone in the case of a negativetest, then travel to the nitrocellulose strip and travel through andpass the test result zone and the control zone of the device. If theanalyte of interest is present in the sample, the analyte-labeledreagent complex binds to the immobilized reagent of the test result zoneforming a detectable line, and if the analyte of interest is not presentin the sample, no analyte labeled reagent complex is formed andtherefore no binding occurs at the test result zone. Whether or not theanalyte of interest is present in the sample to form a complex, thelabeled reagent binds to the immobilized reagent of the control zoneforming a detectable line indicating that the test has run tocompletion. Any excess liquid sample, after the testing has run tocompletion, can be absorbed at the distal sink of the accused device.

Provided is a method of detecting the presence of or measuring theamount of an analyte of interest in sample including the steps ofproviding a sample suspected of containing an analyte of interest. Thesample can be any sample, such as for example, biological fluids orenvironmental samples. The method may also include providing aimmunochromatographic test strip comprising one or more biologicallyactive molecules and one or more non-sugar organic compatible solutes.

The immunochromatographic test strip is preferably associated with animmunoassay like the devices described above for detecting the presenceof or measuring the amount of an analyte of interest in a sample. Themethod of the present invention also includes the step of contacting thesample suspected of containing the analyte of interest with theimmunochromatographic test strip.

The method may also include detecting the presence of or measuring theamount of the analyte of interest The detection of the presence ormeasurement of the amount of analyte of interest can be done by anysuitable method for that purpose, such as for example, use of labeledantibodies or antibody fragments wherein the label includes one or moreof, for example, latex, dye, gold sols, radioactive, or fluorescentlabels.

An important aspect of the present invention includes the ability of theone or more non-sugar organic compatible solutes to protect and preservethe activity of the one or more biologically active molecules in thedried and reconstituted state, e.g. particularly in a conjugate pad orother zone comprising a mobilizable, dried binding agent, but alsobinding agents in a detection zone of any of the above-describeddevices. The one or more non-sugar organic compatible solutes of thepresent invention may be selected from, for example, amino acids such asglycine, alanine, serine, proline, sodium glutamate, lysine, aminobutricacid, taurine, amines such as betaine, sarcosine, trimethylamineN-oxide, or osmolytes or other compatible solutes such as 2-naphthol,ectoine, hydroxyectoine, glycerine, deoxyribonucleic acid, adenosinemonophosphate, PCR primers, spermine, spermidine, Brij5 8,polyethyleneimine, polyhystidine, polythymadilic acid,acetyl-L-camitine, L-citruline, hypotaurine, and ferulic acid.

EXAMPLES

The invention, having been generally described, may be more readilyunderstood by reference to the following examples, which are includedmerely for purposes of illustration of certain aspects and embodimentsof the present invention, and are not intended to limit the invention inany way. All headings are for the convenience of the reader and shouldnot be used to limit the meaning of the text that follows the heading,unless so specified.

Example 1 RSC Capture

According to this example, ICT devices for the detection of RSV antigenswere constructed and used. Capture antibody solutions containingmonoclonal anti-RSV (Binax Inc., Portland Me.) at 1.0 mg/mL and 8% (w/v)Taurine (Sigma, St Louis, Mo.) (protected) or monoclonal anti-RSV (BinaxInc., Portland Me.) at 1.0 mg/mL alone (unprotected) were stripedimmediately after preparation. The capture solutions were striped ontonitrocellulose (HF11 OUBBS, Millipore, Bedford, Mass.) materiallaminated to a polyester film and dried for two minutes at 48° C. Thedevices were then built according to the general design of the deviceusing gold labeled anti-RSV. A subset of the finished devices wasanalyzed the following day (time zero) and the remainder were sealed infoil pouches and stored at 55° C.

Positive (sensitivity, low and moderate) and negative samples for theRSV antibody were applied to the extraction pad of the devices and after15 minutes they were analyzed for detection of the RSV antigens. Initialtesting resulted in a similar response to the positive controls for boththe unstabilized and stabilized striped capture solutions. In all casesthe negative control did not generate a positive signal. After one weekat 55° C. the unprotected devices failed to detect the sensitivitycontrol. By three weeks the low control produced only faint signal forthe unprotected devices, while the protected devices generated aresponse nearly equivalent to that of the protected devices run at weekone. Lastly, after two months the unprotected devices were all butunresponsive to the three positive controls. The protected devicescontinued to detect the RSV antigen in the low and moderate controls.

Example 2 Streptococcus Pneumoniae Capture

According to this example, ICT devices for the detection of S.pneumoniae antigens were constructed and used. Capture antibodysolutions containing affinity purified rabbit anti-Strep pneumoniaeantibody (Binax Inc, Portland Me.) at 1.25 mg/ml and 5% (w/v)Hydroxyectoine (Fluka Production, GmbH) (protected) or affinity purifiedrabbit anti-Strep pneumoniae antibody (Binax Inc, Portland Me.) at 1.25mg/ml alone (unprotected) were striped immediately after preparation.The capture solutions were striped onto nitrocellulose (SR, Millipore,Bedford, Mass.) material laminated to a polyester film and dried for twominutes at 50° C. The devices were then built according to the generaldesign of the device using gold labeled rabbit anti-Strep pneumoniaeantibody. A subset of the finished devices was analyzed the followingday (time zero) and the remainder were sealed in foil pouches and storedat 55° C.

Positive (sensitivity low and moderate) and negative samples for theStrep pneumoniae antibody were applied to the patient swab in the devicefollowed by the addition of the chase solution and after 15 minutes theywere analyzed for detection of the Strep pneumoniae antigens. In allcases the negative control did not generate a positive signal. After oneweek at 55° C. the unprotected devices generated signals half that ofthe devices stabilized with hydroxyectoine. After one month the lowcontrol was not detectable with unprotected devices, while the protecteddevices continued to detect the sensitivity control. After two monthsthe results were similar to that of the one month time point.

Example 3 Filariasis Capture

According to this example, ICT devices for the detection of RSV antigenswere constructed and used. Capture antibody solutions containingmonoclonal anti-D. immitis IgM (Binax Inc, Portland Me.) at 1.5 mg/d and5% (w/v) Spermine (Fluka Production, GmbH) (protected) or monoclonalanti-D. immitis IgM (Binax Inc, Portland Me.) at 1.5 mg/ml alone(unprotected) were striped immediately after preparation. The capturesolutions were striped onto nitrocellulose (HFI lOUBBS, Millipore,Bedford, Mass.) material laminated to a polyester film and dried for twominutes at 50° C. The devices were then built according to the generaldesign of the device using gold labeled anti-D. immitis. A subset of thefinished devices was analyzed shortly thereafter (time zero) and theremainder were sealed in foil pouches and stored at 55° C.

Positive (low and moderate) sera and negative whole blood samples forthe D. immitis antibody were applied to the extraction pad of thedevices and after 15 minutes they were analyzed for detection of the D.immitis antigens. Initial testing resulted in a similar response to thepositive controls for both the unstabilized and stabilized stripedcapture solutions. In all cases the negative control did not generate apositive signal. After one week at 55° C. the unprotected devices failedto detect the low control while the stabilized devices produced a signalequivalent to the initial testing. After one month the unprotecteddevices did not detect either the low or the moderate positive control,while the protected devices detected both the low and moderate positivecontrol. After two months the unprotected devices continued to beunresponsive to the positive controls. The protected devices detectedthe D. immitis antigen in the moderate control.

Example 4 RSV Capture

According to this example, ICT devices for the detection of RSV antigenswere constructed and used. Capture antibody solutions containingmonoclonal anti-RSV (Binax Inc, Portland Me.) at 1.0 mg/mL and 1.25%(w/v) Adenosine S′monophosphate (Sigma, St Louis, Mo.) (protected) wasstriped immediately after preparation. The capture solution was stripedonto nitrocellulose (HFI lOUBBS, Millipore, Bedford, Mass.) materiallaminated to a polyester film and dried for two minutes at 48° C. Thedevices were then built according to the general design of the deviceusing gold labeled anti-RSV. A subset of the finished devices wasanalyzed the following day (time zero) and the remainder were sealed infoil pouches and stored at 55° C.

Positive (sensitivity, low and moderate) and negative samples for theRSV antibody were applied to the extraction pad of the devices and after15 minutes they were analyzed for detection of the RSV antigens. Initialtesting of the devices with the stabilized RSV antibody detected the RSVantigen in each of the three positive controls, sensitivity, low andmoderate. In all cases the negative control did not generate a positivesignal. After one week at 55° C. the signal generated by each of thecontrol type was the same as that of the initial testing. The devicescontinued to detect the RSV antigen in the sensitivity, low and moderatecontrols after two months at 55° C.

Example 5 Immunochromatographic Test for Malaria

Non-sugar organic-compatible solutes were used to stabilize antibody onnitrocellulose and gold. A concentration of 0.1% polyhistidine was ableto stabilize the aldolase recombinant. A sample stored at 30° C. wasshown to have stability for a minimum of 29 days when it was previouslystable for only 2 days.

Example 6 Combination of 2% W/V of Ecotine L-Histidine, Taurine andL-Threonine Confers Stability to Conjugate Pad Drying Buffer w/o Sucrose

A 5× conjugate drying buffer was prepared following the SOP formulationexcept that the SOP 25% w/v sucrose was excluded. A combination ofcontrol, anti P. falciparum and anti P. vivax gold particles wereadjusted to 1× by addition of 5× drying buffer and brought to 2% w/veach of ecotine L-histidine, taurine and L-threonine. SOP conjugate padmatrix was striped, dried and stored in desiccated vials at both 4° C.and 55° C. for 19 months and 11 days. Vials were removed to RT andallowed to equilibrate and pads from each temperature were inserted intomalaria urine dipstick format devices. The same combination ofrecominant P. vivax antigen and tissue culture supernatant P. falciparumantigen was run on one of each device. Equivalent sensitivity wasobserved for conjugate pad stored at both 4° C. and 55 ° C. despite theabsence of sucrose in the drying pad buffer formulation.

Example 7

4% W/V of L-Histidine Alone does not Confer Stability to Conjugate PadDrying Buffer w/o Sucrose

5× conjugate drying buffer was prepared following the SOP formulationexcept that the SOP 25% w/v sucrose was excluded. A combination ofcontrol, anti P. falciparum and anti P. vivax gold particles wereadjusted to 1× by addition of 5× drying buffer and brought to 4% w/v.SOP conjugate pad matrix was striped, dried and stored in desiccatedvials at both 4° C. and 55° C. for 19 months and 11 days. Vials wereremoved to RT and allowed to equilibrate and pads from each temperaturewere inserted into malaria urine dipstick format devices. The samecombination of recominant P. vivax antigen and tissue culturesupernatant P. falciparum antigen was run on one of each device. Muchhigher sensitivity was observed for conjugate pad stored at 4° C. thanfor pad stored at 55° C. Additionally, a significant quantity ofconjugate was retained in the conjugate pad matrix of the 55° C.material after the run.

Equivalents

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to theembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims.

1. A method, comprising: a) depositing one or more biologically activemolecules onto a surface; b) depositing one or more non-sugar organiccompatible solutes onto the surface; and c) drying the deposited one ormore biologically active molecules and the deposited one or morenon-sugar organic compatible solutes on the surface, wherein the one ormore non-sugar organic compatible solutes preserves the activity of theone or more biologically active molecules in the dried and reconstitutedstate.
 2. The method of claim 1, wherein the one or more biologicallyactive molecules and the one or more non-sugar organic compatiblesolutes are deposited onto said surface concurrently.
 3. The method ofclaim 2, wherein each of the one or more biologically active moleculesare selected from the group consisting of: an antibody, a labeledantibody, an antibody fragment and a labeled antibody fragment, and theone or more non-sugar organic compatible solutes comprise single aminoacids.
 4. The method of claim 2, wherein the one or more biologicallyactive molecules are selected from the group consisting of: an antibody,a labeled antibody, an antibody fragment and a labeled antibodyfragment, and the one or more non-sugar organic compatible solutescomprise single amino acid chains of about 1 to about 10 amino acids. 5.The method of claim 3, wherein each of the one or more biologicallyactive molecules are selected from the group consisting of: animmobilized antibody, an immobilized antibody fragment, a mobilizablelabeled antibody and a mobilizable labeled antibody fragment, and thesurface comprises a portion of a test strip.
 6. The method of claim 5,wherein the test strip is immunochromatographic.
 7. The method of claim5, wherein the portion of the test strip is a conjugate pad.
 8. Themethod of claim 5, wherein the portion of the test strip is a detectionzone.
 9. The method of claim 5, wherein said immunochromatographic stripcomprises a lateral flow device.
 10. The method of claim 2, wherein saidone or more biologically active molecules are selected from the groupconsisting of: an antibody, a labeled antibody, an antibody fragment anda labeled antibody fragment, and said one or more non-sugar organiccompatible solutes comprise one or more amines.
 11. The method of claim2, wherein said one or more biologically active molecules are selectedfrom the group consisting of: an antibody, a labeled antibody, anantibody fragment and a labeled antibody fragment, and said one or morenon-sugar organic compatible solutes comprise osmolytes or othercompatible solutes.
 12. A method, comprising: a) providing a sample; b)providing a test strip with at least a portion comprising one or morebiologically active molecules and one or more non-sugar organiccompatible solutes, wherein said one or more non-sugar organiccompatible solutes protects and preserves the activity of said one ormore biologically active molecules in the dried and reconstituted state;c) contacting said with said test strip; and d) detecting the presenceof or measuring the amount of an analyte in the sample, wherein said oneor more non-sugar organic compatible solutes protects and preserves theactivity of said one or more biologically active molecules in the driedand reconstituted state.
 13. The method of claim 12, wherein the portionof the test strip is a conjugate pad.
 14. The method of claim 12,wherein the portion of the test strip is a detection zone.
 15. Themethod of claim 12, wherein the test strip is immunochromatographic. 16.The method of claim 12, wherein the test strip comprises a lateral flowdevice.
 17. The method of claim 12, wherein said one or more non-sugarorganic compatible solutes are each selected from the group consistingof: glycine, alanine, serine, proline, sodium glutamate, lysine,aminobutric acid, taurine, amines, betaine, sarcosine, trimethylamineN-oxide, 2-Naphthol, ectoine, hydroxyectoine, glycerine,deoxyribonucleic acid, adenosine monophosphate, PCR primers, spermine,spermidine, BnJ5 8, polyethyleneimine, polyhystidine, polythymadilicacid, acetyl-L-carnitine, and L-citruline.
 18. A test strip defining aflow path and comprising: a porous member comprising, in dry form: alabeled binding reagent configured to bind an analyte, and at least onenon-sugar organic compatible solute, a porous strip separate from and influid contact with the porous member, the porous strip comprising adetection zone, and a binding reagent disposed along the flow path andconfigured to bind the analyte and/or a complex of the analyte and thelabeled binding reagent: wherein, the binding reagent is configured tocapture the analyte and/or the complex of the analyte and the labeledbinding reagent in the detection zone upon the mobilization of thelabeled binding reagent by a liquid comprising the analyte, and thenon-sugar organic compatible solute is selected from the groupconsisting of glycine, alanine, serine, proline, sodium glutamate,lysine, aminobutric acid, taurine, amines, betaine, sarcosine,trimethylamine N-oxide, 2-Naphthol, ectoine, hydroxyectoine, glycerine,deoxyribonucleic acid, adenosine monophosphate, PCR primers, spermine,spermidine, BnJ5 8, polyethyleneimine, polyhystidine, polythymadilicacid, acetyl-L-carnitine, and L-citruline.